An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)

The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the muta...

Full description

Bibliographic Details
Main Authors: Abdul Hamid, Azzmer Azzar, Tengku Abdul Hamid, Tengku Haziyamin, Abdul Wahab, Roswanira, Omar, Mohd. Shahir Shamsir, Huyop, Fahrul
Format: Article
Language:English
Published: Penerbit UTM Press 2015
Subjects:
Online Access:http://eprints.utm.my/57794/1/RoswaniraAbdulWahab2015_AnS188VMutationAltersSubstrateSpecificity.pdf
_version_ 1796860380440428544
author Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
author_facet Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
author_sort Abdul Hamid, Azzmer Azzar
collection ePrints
description The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves substrate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2-chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by Km. This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies.
first_indexed 2024-03-05T19:40:30Z
format Article
id utm.eprints-57794
institution Universiti Teknologi Malaysia - ePrints
language English
last_indexed 2024-03-05T19:40:30Z
publishDate 2015
publisher Penerbit UTM Press
record_format dspace
spelling utm.eprints-577942021-08-04T05:07:17Z http://eprints.utm.my/57794/ An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE) Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Omar, Mohd. Shahir Shamsir Huyop, Fahrul QD Chemistry The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated compounds but is ineffective against β-halogenated compounds such as 3-chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves substrate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2-chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by Km. This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies. Penerbit UTM Press 2015 Article PeerReviewed application/pdf en http://eprints.utm.my/57794/1/RoswaniraAbdulWahab2015_AnS188VMutationAltersSubstrateSpecificity.pdf Abdul Hamid, Azzmer Azzar and Tengku Abdul Hamid, Tengku Haziyamin and Abdul Wahab, Roswanira and Omar, Mohd. Shahir Shamsir and Huyop, Fahrul (2015) An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE). Plos One, 10 (3). pp. 1-21. ISSN 1932-6203 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0121687
spellingShingle QD Chemistry
Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title_full An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title_fullStr An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title_full_unstemmed An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title_short An S188V Mutation Alters Substrate Specificity of Non- Stereospecific alpha-Haloalkanoic Acid Dehalogenase E (DehE)
title_sort s188v mutation alters substrate specificity of non stereospecific alpha haloalkanoic acid dehalogenase e dehe
topic QD Chemistry
url http://eprints.utm.my/57794/1/RoswaniraAbdulWahab2015_AnS188VMutationAltersSubstrateSpecificity.pdf
work_keys_str_mv AT abdulhamidazzmerazzar ans188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT tengkuabdulhamidtengkuhaziyamin ans188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT abdulwahabroswanira ans188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT omarmohdshahirshamsir ans188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT huyopfahrul ans188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT abdulhamidazzmerazzar s188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT tengkuabdulhamidtengkuhaziyamin s188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT abdulwahabroswanira s188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT omarmohdshahirshamsir s188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe
AT huyopfahrul s188vmutationalterssubstratespecificityofnonstereospecificalphahaloalkanoicaciddehalogenaseedehe